Identifikacija mehanizma nastanka stresnih granul v celicah po obdelavi s hladno atmosfersko plazmo

Karlek, Anže (avtor), Rogelj, Boris (mentor)

Hladna atmosferska plazma (HAP) je ioniziran plin s približno sobno temperaturo. HAP izkazuje različne fizikalno-kemične lastnosti, saj tekom pridobivanja HAP nastajajo s reaktivne spojine, nevtralni delci, elektromagnetno polje in UV sevanje. HAP je mogoče uporabiti v različne namene, posvetili pa smo se uporabi v medicini. Do sedaj je znano, da HAP povzroča nastanek stresnih granul (SG) po od eIF2α-odvisni poti, vendar točen mehanizem aktivacije integriranega celične odziva (ISR) s strani HAP še ni poznan. Glede na dosedanje poznavanje delovanja HAP vemo, da se ob delovanju HAP tvorijo kisikove in dušikove reaktivne spojine. Z dodatkom znanega antioksidanta kverceta k celični liniji SH-SY5Y Flp-In mScarletI-G3BP1-Myc, smo preverili vpliv oksidativnega stresa na tvorbo SG ob delovanju HAP. Pri tem smo predvidevali, da kvercetin zavira aktivacijo kinaze HRI, ki je odgovorna za nastanek SG ob prisotnosti oksidativnega stresa. Nastanek in lastnosti SG smo opazovali v celični liniji SH-SY5Y Flp-In mScarletI-G3BP1-Myc, ki ima na protin G3BP1 vezan rdeči fluorescenčni protein mScarletI. Protein G3BP1 je eden izmed proteinov v SG, zato se ob delovanju stresa rekrutira v SG, kar opazujemo s fluorescenčno konfokalnim mikroskopom. HAP smo pridobivali s plazemsko šobo pri konstantnem dotoku argona. Pokazali smo, da z dodatek kvercetina k celicam zmanjša delež celic s stresnimi granulami, hkrati pa vpliva na stresni stimulus HAP. Tako smo potrdili vpletenost kisikovih in dušikovih reaktivnih spojin v tvorbi SG. Vpliv kvercetina na fosforilacijo eIF2α nismo dokazali.

• Jezik: Slovenski jezik
• Ključne besede: hladna atmosferska plazma, stresne granule, kvercetin, natrijev arzenit, celični stres, oksidativni stres, eIF2α
• Vrsta gradiva: Magistrsko delo/naloga
• Tipologija: 2.09 - Magistrsko delo
• Organizacija: FKKT - Fakulteta za kemijo in kemijsko tehnologijo
• Leto izida: 2023
• PID: 20.500.12556/RUL-150938
• COBISS.SI-ID: 173210627
• Datum objave v RUL: 26.09.2023

KARLEK, Anže, 2023, Identifikacija mehanizma nastanka stresnih granul v celicah po obdelavi s hladno atmosfersko plazmo [na spletu]. Magistrsko delo. [Dostopano 4 maj 2025]. Pridobljeno s: https://repozitorij.uni-lj.si/IzpisGradiva.php?lang=slv&id=150938

Sekundarni jezik

• Jezik: Angleški jezik
• Naslov: Identification of the stress granule formation mechanism induced in cells upon cold atmospheric plasma treatment

Izvleček

Cold atmospheric plasma (CAP) is a near-room temperature ionized gas that is widely used in biomedicine. CAP cause physicochemical effects, as various processes take place during the generation of CAP, such as reactive compounds, neutral particles, electromagnetic fields and UV radiation. It is well established that CAP causes the formation of stress granules via the eIF2α-phosphorylation dependent pathway. However, it is unknown which of the integrated stress response related kinases is responsible for the phosphorylation of eIF2α. We can assume that CAP is responsible for oxidative stress due to the generation of reactive oxygen and nitrogen species by the application of CAP. Quercetin is a known antioxidant, so we assumed that quercetin may inhibit the activation of heme-regulated inhibitor kinase, which is activated during oxidative stress. We used the cell line SH-SY5Y Flp-In mScarletI-G3BP1-Myc, in which the fluorescent protein mScarletI is fused to one of the core proteins of stress granules, G3BP1. The source of CAP was a plasma jet with constant argon flow. We demonstrated that quercetin significantly reduced the number of cells with stress granules when incubated for one hour before treatment with CAP. Quercetin also acts on strength of stress stimuli caused by CAP and on affects stress granule morphology. We did not detect any effect of quercetin on eIF2α phosphorylation.

• Ključne besede: cold atmospheric plasma, stress granules, quercetin, sodium arsenite, cell stress, oxidative stress, eIF2α